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The human pS2 gene is specifically expressed is a subclass of estrogen receptor containing human breast cancer cells. In the MCF7 cell line, its induction by estradiol is a primary transcriptional event. The exact location of its estrogen responsive element has been determined using a chimeric recombinant transfected into HeLa cells and a transient expression assay. In this study we found, using electrophoretic mobility shift experiments, that in HeLa cells the estrogen responsive element (ERE) of the pS2 gene is recognized by a methylation sensitive DNA binding protein (MSDBP) different from the estrogen receptor. Competition experiments have shown that the binding of this protein requires at least one CpG in the center of the palindromic sequence and that imperfect palindromic sequences are also recognized. Although the presence of CpG is necessary, CpG-rich oligonucleotides, containing consensus sequences for Sp1 or AP2, do not interfere with its binding to the pS2 oligonucleotide, indicating that the ERE sequence itself participates in the specificity of its binding. This protein binds the pS2 sequence with a relatively high affinity (apparent Kd = 10(-10) M) and its binding is strongly reduced by the methylation of the cytosines at CpG sites. UV cross-linking experiments and peptide mapping indicate that this protein has an apparent molecular weight of 46 kDa and is present in several cell lines, including non-human cell lines. Taken together, these data suggest that this protein might have a potential role in regulating gene activity or in chromatin structure of some genes possessing an ERE.

The Estrogen Responsive Element of the pS2 Gene Is Recognized by a Methylation Sensitive DNA Binding Protein